I've cultured primary microglia from neonates in 24-well plates, and typically I lift via scraping and put the cells straight into flow tubes for analysis. However, I would like to do additional staining this time, so I need to lift the microglia, transfer to a v-bottom plate, and do the staining in the plate. I've tried centrifuging the microglia in the past, but I've always lost all of my cells. Does anybody have any recommendations for lifting or time/speed for centrifugation in a 96well v bottom plate as to not lose the cells after staining? I don't want to stain the cells in 24 well plates because of the amount of antibody I would need to use.
Thanks!
From the recent preprint that uses our DendroTEK dPGA surface coating.
"Flasks were first sealed with Parafilm (#52858-000, Bemis) and shaken for 1 h at 37°C, at 150 rpm in an orbital shaking incubator with a rotational diameter of 3 cm (Jeio Tech, model SI600), to detach the microglia, but not the OPCs or astrocytes, which remain attached to the bottom of the flask. Detached microglia were removed with the culture medium."
Source: https://www.biorxiv.org/content/10.1101/2025.06.23.661031v1.abstract